Cationic hydrogels prepared from regioselectively azidated (1→3)-α-d-glucan via crosslinking and amination: Physical and adsorption properties.

Cationic hydrogels with amino groups were successfully prepared using (1→3)-α-d-glucan synthesized by glucosyltransferase J (GtfJ) cloned from Streptococcus salivarius through a three-step reaction: (i) Azido groups were regioselectively introduced at the C6 position of (1→3)-α-d-glucan by a bromination-azidation process (degree of substitution 0.94), (ii) Azido groups were partially crosslinked with 1,8-nonadiyne via a copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction, (iii) Azido groups that were unused for crosslinking were reduced to amino groups by sodium borohydride (NaBH4). The introduction of amino groups was confirmed quantitatively and qualitatively by elemental, Fourier transform infrared (FT-IR), and nuclear magnetic resonance (NMR) analyses. These cationic hydrogels showed a specific adsorption ability for bovine serum albumin (BSA) over a wide pH range of 4.5-8.0 due to their high pH values at the point of zero charge (pHpzc 8.80-8.92).

Enzymatic polymerization derived engineered polysaccharides as reinforcing fillers of ethylene vinyl acetate composites.

A novel monomer based, controlled enzymatic polymerization was employed to produce an engineered alpha-1,3 glucan polysaccharide. The structure and material properties of the engineered polysaccharide were characterized using various techniques. The use of such engineered polysaccharide as a reinforcing filler of polymers was evaluated using model polymers. For this, the alpha-1,3 glucan was incorporated into ethylene vinyl acetate co-polymer (EVA) matrices with vinyl acetate content of 32% and 40% via a melt processing fabrication process. Various mechanical and rheological properties of the fabricated composites were evaluated. The effect of vinyl acetate content of the EVA resin on the interaction with alpha-1,3 glucan that result in various performances attributes was also investigated and reported. The incorporation of alpha-1,3 glucan in these EVA composites resulted in the improvement of key composite properties, such as toughness, modulus, wear resistance, and hardness showing the reinforcing potential of these engineered polysaccharides.

Rapid urease test (RUT) for evaluation of urease activity in oral bacteria in vitro and in supragingival dental plaque ex vivo.

BACKGROUND: Urease is an enzyme produced by plaque bacteria hydrolysing urea from saliva and gingival exudate into ammonia in order to regulate the pH in the dental biofilm. The aim of this study was to assess the urease activity among oral bacterial species by using the rapid urease test (RUT) in a micro-plate format and to examine whether this test could be used for measuring the urease activity in site-specific supragingival dental plaque samples ex vivo. METHODS: The RUT test is based on 2% urea in peptone broth solution and with phenol red at pH 6.0. Oral bacterial species were tested for their urease activity using 100 µl of RUT test solution in the well of a micro-plate to which a 1 µl amount of cells collected after growth on blood agar plates or in broth, were added. The color change was determined after 15, 30 min, and 1 and 2 h. The reaction was graded in a 4-graded scale (none, weak, medium, strong). Ex vivo evaluation of dental plaque urease activity was tested in supragingival 1 µl plaque samples collected from 4 interproximal sites of front teeth and molars in 18 adult volunteers. The color reaction was read after 1 h in room temperature and scored as in the in vitro test. RESULTS: The strongest activity was registered for Staphylococcus epidermidis, Helicobacter pylori, Campylobacter ureolyticus and some strains of Haemophilus parainfluenzae, while known ureolytic species such as Streptococcus salivarius and Actinomyces naeslundii showed a weaker, variable and strain-dependent activity. Temperature had minor influence on the RUT reaction. The interproximal supragingival dental plaque between the lower central incisors (site 31/41) showed significantly higher scores compared to between the upper central incisors (site 11/21), between the upper left first molar and second premolar (site 26/25) and between the lower right second premolar and molar (site 45/46). CONCLUSION: The rapid urease test (RUT) in a micro-plate format can be used as a simple and rapid method to test urease activity in bacterial strains in vitro and as a chair-side method for testing urease activity in site-specific supragingival plaque samples ex vivo.

Characterization of crystalline linear (1→3)-α-d-glucan synthesized in vitro.

We investigated the crystal structure and molecular arrangement of the linear (1→3)-α-d-glucan synthesized by glucosyltransferase GtfJ cloned from Streptococcus salivarius using sucrose as a substrate. The synthetic products had two morphologies: wavy fibril-like crystals as major and thin lamellae as minor products. Their structures were analyzed using electron microdiffraction, synchrotron X-ray powder diffraction, and solid-state 13C NMR spectroscopy. The fibrils and lamellae had the same allomorphic form but different molecular arrangements. The wet crystals were in a hydrated form, which converted into an anhydrous form with a significant decrease in crystallinity on drying. The hydrated and anhydrous forms had an extended-chain conformation with 2/1 helix, and the hydrated form was estimated to contain one water molecule per glucose residue. The long glucan chains were folded in the fibril crystals, while the short, extended chains were arranged perpendicular to the base plane of the lamellae.

Salivaricin E and abundant dextranase activity may contribute to the anti-cariogenic potential of the probiotic candidate Streptococcus salivarius JH.

Dental caries is an infectious disease that is continuing to increase in prevalence, reducing the quality of life for millions worldwide as well as causing considerable expense, with an estimated US$108 billion spent on dental care in the USA each year. Oral probiotics are now being investigated to determine whether they could play a role in the prevention and treatment of this disease. Streptococcus salivarius strain JH is a potential probiotic candidate that produces multiple proteinaceous antimicrobials (bacteriocins), the inhibitory spectrum of which includes Streptococcus mutans, one of the principal causative agents of dental caries. The genome of strain JH has previously been shown to contain the biosynthetic loci for the bacteriocins salivaricin A3, streptin and streptococcin SA-FF22. Here we show that strain JH also produces salivaricin E, a 32 aa lantibiotic with a mass of 3565.9 Da, which is responsible for the inhibition of S. mutans growth. In addition, strain JH was shown to produce dextranase, an enzyme that hydrolyses (1 → 6)-α-D-glucosidic linkages, at levels higher than any other S. salivarius tested. In vitro testing showed that partial hydrolysis of the exopolymeric substances of S. mutans, using strain JH dextranase, improved the anti-S. mutans inhibitory activity of the lytic bacteriocin, zoocin A. The multiple bacteriocin and dextranase activities of strain JH support its candidature for development as an oral probiotic.